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Mesenchymal stem cells isolated from rats are frequently used fortissue engineering research. However, considerable differences have been identifiedbetween rat mesenchymal stem cells and those derived from humans, and no defined panelof markers currently exists for the isolation of these cells. The aim of this study wasto examine the effects of cell sorting for CD29+/CD90+ cells from rat adipose and bonemarrow tissues on their differentiation and expression of stem cell–associated genes.Flow cytometry showed 66% and 78% CD29+/CD90+ positivity within passage 1 of adipose andbone marrow cultures, respectively. CD29+/CD90+ cells showed a reduction in bothosteogenic and adipogenic differentiation when compared with unsorted cells, asdetermined by alizarin red and Oil Red-O staining, respectively. These findings couldnot entirely be explained by fluorescence-activated cell sorting–induced cell injury assort recovery was only modestly affected in adipose-derived cells. Maintaining cells influorescence-activated cell sorting buffer did not affect adipose-derived cellviability, but a significant (p < 0.05) reduction was found in bone marrow–derivedcell viability. Additionally, CD29+/CD90+ selection was associated with a significantdecrease in the expression of Lin28, Sox2, Nanog and CD73 in adipose-derived cellcultures, whereas differences in stem cell–associated gene expression were not observedin sorted bone marrow–derived cell cultures. In summary, this study demonstrated thatfluorescence-activated cell sorting had differential effects on adipose-derived cellsand bone marrow–derived cells, and both CD29+/CD90+ cells displayed a significantlyreduced capacity for osteogenic/adipogenic differentiation. In conclusion, we identifythat maintaining heterogeneity within the mesenchymal stem cell population may beimportant for optimal differentiation

Isolation of adipose and bone marrow mesenchymal stem cells using CD29 and CD90 modifies their capacity for osteogenic and adipogenic differentiation